Modeling the Impact of Agricultural Mitigation Measures on the Spread of Sharka Disease in Sweet Cherry Orchards.

Sharka is a disease affecting stone fruit trees. It is caused by the Plum pox virus (PPV), with Myzus persicae being one of the most efficient aphid species in transmitting it within and among Prunus orchards. Other agricultural management strategies are also responsible for the spread of disease among trees, such as grafting and pruning. We present a mathematical model of impulsive differential equations to represent the dynamics of Sharka disease in the tree and vector population. We consider three transmission routes: grafting, pruning, and through aphid vectors. Grafting, pruning, and vector control occur as pulses at specific instants. Within the model, human risk perception towards disease influences these agricultural management strategies. Model results show that grafting with infected biological material has a significant impact on the spread of the disease. In addition, detecting infectious symptomatic and asymptomatic trees in the short term is critical to reduce disease spread. Furthermore, vector control to prevent aphid movement between trees is crucial for disease mitigation, as well as implementing awareness campaigns for Sharka disease in agricultural communities that provide a long-term impact on responsible pruning, grafting, and vector control.

A fast and efficient protocol for small RNA extraction in Japanese plum and other Prunus species

Background: Small ribonucleic acids represent an important repertoire of mobile molecules that exert key roles in several cell processes including antiviral defense. Small RNA based repertoire includes both small interfering RNA (siRNA) and microRNA (miRNA) molecules. In the Prunus genus, sharka disease, caused by the Plum pox virus (PPV), first occurred on European plum (Prunus domestica) and then spread over among all species in this genus and thus classified as quarantine pathogen. Next-generation sequencing (NGS) was used for the study of siRNA/miRNA molecules; however, NGS relies on adequate extraction protocols. Currently, knowledge of PPV-Prunus interactions in terms of siRNA populations and miRNA species is still scarce, and siRNA/miRNA extraction protocols are limited to species such as peach, almond, and sweet cherry. Results: We describe a reliable procedure for siRNA/miRNA purification from Prunus salicina trees, in which previously used protocols did not allow adequate purification. The procedure was based on a combination of commercially available RNA purification kits and specific steps that yielded high quality purifications. The resulting molecules were adequate for library construction and NGS, leading to the development of a pipeline for analysis of both siRNAs and miRNAs in the PPV­P. salicina interactions. Results showed that PPV infection led to altered siRNA profiles in Japanese plum as characterized by decreased 24-nt and increased 21- and 22-nt siRNAs. Infections showed miR164 and miR160 generation and increased miR166, miR171, miR168, miR319, miR157, and miR159. Conclusion: We propose this protocol as a reliable and reproducible small RNA isolation procedure for P. salicina and other Prunus species.